Wheat bran proteolysis peptide / Chinese wolfberry fruit composite beverage

Abstract

The invention relates to the technical field of food processing, and discloses a wheat bran proteolysis peptide / Chinese wolfberry fruit composite beverage. The wheat bran protein enzymolysis peptide / Chinese wolfberry fruit compound beverage provided by the invention is prepared from the following raw materials in percentage by mass: 15 to 35 percent of wheat bran protein enzymolysis peptide, 1.4 to 2.2 percent of Chinese wolfberry fruit juice, 4 to 8 percent of stevioside, 0.2 to 0.4 percent of citric acid, 0.1 percent of sodium carboxymethyl cellulose, 0.08 percent of xanthan gum and the balance of water. The wheat bran proteolysis peptide / fructus lycii compound beverage provided by the invention has a relatively high sensory evaluation score and relatively good in-vitro blood fat reducing activity.

Description

Technical Field

[0001] The invention relates to the technical field of food processing, in particular to a wheat gluten enzymatic hydrolyzed peptide / wolfberry composite beverage. Background Art

[0002] In recent years, with the continuous improvement of people's living standards, eating habits and lifestyles have also changed significantly, which has led to an increasing incidence of hyperlipidemia year by year. Hyperlipidemia has a high incidence and is very harmful, especially after entering middle-aged and elderly people. Hyperlipidemia is more likely to induce atherosclerosis, thereby causing a series of cardiovascular and cerebrovascular diseases, such as coronary heart disease, stroke, etc., which can even be life-threatening in severe cases. Therefore, hyperlipidemia has become a hot topic in people's health.

[0003] Functional drinks, as a special kind of beverage, usually contain a variety of vitamins, minerals, amino acids and other functional ingredients. Their purpose is to meet the nutritional needs of the human body and regulate the physiological functions of the human body to a certain extent. Therefore, if functional drinks are made to have a certain lipid-lowering activity, people can reduce the risk of hyperlipidemia by drinking these functional drinks with a certain lipid-lowering activity. Therefore, providing a functional drink with good lipid-lowering activity has become a technical problem that needs to be urgently solved by technical personnel in this field. Summary of the invention

[0004] The purpose of the present invention is to provide a gluten enzymatic peptide / wolfberry composite beverage, which has a higher sensory evaluation score and better in vitro blood lipid lowering activity.

[0005] To achieve the above object, the present invention provides the following technical solutions:

[0006] One of the technical solutions of the present invention:

[0007] A wheat gluten enzymatic peptide / wolfberry composite beverage, comprising the following raw materials in percentage by weight:

[0008] 15% to 35% of wheat gluten enzymatic peptides, 1.4% to 2.2% of wolfberry juice, 4% to 8% of stevioside, 0.2% to 0.4% of citric acid, 0.1% of sodium carboxymethyl cellulose, 0.08% of xanthan gum, and the balance of water.

[0009] Furthermore, the method for preparing the wheat bran enzymatic hydrolysis peptide comprises the following steps:

[0010] 1) Using wheat bran as raw material, extracting wheat bran protein by alkali dissolution and acid precipitation method;

[0011] 2) Using a composite protease to enzymatically hydrolyze the gluten protein obtained in step 1) to obtain the gluten enzymatic hydrolyzed peptide.

[0012] Furthermore, in step 1), the alkali dissolution and acid precipitation method is specifically as follows: the wheat bran is crushed through a 60-mesh sieve and defatted with n-hexane to obtain defatted wheat bran; the defatted wheat bran is added to water according to a solid-liquid ratio of defatted wheat bran to water of 1:10, and the pH of the reaction system is adjusted to 10.0 with 1 mol / L NaOH, stirred at 50°C for 1 hour, and centrifuged at 4500 rpm for 10 minutes to obtain a supernatant; the pH of the supernatant is adjusted to 4.0 with 1 mol / L HCl, the precipitate is recovered, and then the pH of the supernatant is adjusted to 7.0 with 1 mol / L NaOH, and frozen to obtain the wheat bran protein.

[0013] Furthermore, in step 2), the enzymolysis is specifically as follows: dissolving the gluten protein obtained in step 1) in water according to a mass volume ratio of gluten protein to water of 2g:100mL, adding a composite protease according to an enzyme addition amount of 14000U / g, hydrolyzing for 3h at 45°C and pH=7.8, and then heating at 95°C for 10min to inactivate the composite protease to obtain a gluten protein hydrolysate; controlling the amount of the gluten protein hydrolysate added to be 2 / 3 of the volume of the dialysis bag, adding the gluten protein hydrolysate to the dialysis bag washed with distilled water, and sealing the bag; placing the dialysis bag containing the gluten protein hydrolysate in distilled water at 4°C, magnetically stirring for 24h, to obtain a desalted gluten protein hydrolysate; first placing the desalted gluten protein hydrolysate in a -20°C refrigerator, freezing for 12h, then placing it in a -80°C refrigerator, freezing for 24h, and finally freeze-drying for 72h.

[0014] Furthermore, the composite protease contains Bacillus subtilis, and its enzyme activity is 120U / mg.

[0015] Furthermore, the method for preparing wolfberry juice comprises the following steps:

[0016] The wolfberries were soaked in distilled water at room temperature for 4 hours, and then the soaked wolfberries were mixed with water at a mass volume ratio of 1 g: 2 mL, and pulped to obtain the wolfberry juice.

[0017] The second technical solution of the present invention:

[0018] The method for preparing the above-mentioned wheat gluten enzymatic peptide / wolfberry composite beverage comprises the following steps:

[0019] 1) Weigh each raw material according to the above mass percentage;

[0020] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] The sensory evaluation score of the wheat gluten enzymatic hydrolyzed peptide / wolfberry composite beverage provided by the present invention can reach 93.50±1.54, which has a relatively high sensory evaluation score;

[0023] The wheat gluten enzymatic peptide / wolfberry composite beverage provided by the invention has an inhibition rate of 81.44±3.30% on cholesterol micelle solubility, 57.47±2.89% on cholesterol esterase and 58.04±3.47% on pancreatic lipase, and has good in vitro lipid-lowering activity. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Various other advantages and benefits will become apparent to those of ordinary skill in the art by reading the detailed description of the preferred embodiments below. The accompanying drawings are only for the purpose of illustrating the preferred embodiments and are not to be considered as limiting the present invention. Moreover, the same reference symbols are used throughout the accompanying drawings to represent the same components. In the accompanying drawings:

[0025] Figure 1 The sensory evaluation results of the wheat gluten enzymatic peptide / wolfberry composite beverage prepared in Examples 1 to 5;

[0026] Figure 2 The sensory evaluation results of the wheat gluten enzymatic peptide / wolfberry composite beverage prepared in Examples 6 to 10;

[0027] Figure 3 The sensory evaluation results of the wheat gluten enzymatic peptide / wolfberry composite beverage prepared in Examples 11 to 15;

[0028] Figure 4 The sensory evaluation results of the wheat gluten enzymatic peptide / wolfberry composite beverage prepared in Examples 16 to 20;

[0029] Figure 5 The results of in vitro lipid-lowering activity test of the wheat bran enzymatic peptide / wolfberry composite beverage of the present invention are shown in Figure 1, wherein A is the cholesterol micelle solubility inhibition rate, B is the cholesterol esterase inhibition rate, and C is the pancreatic lipase inhibition rate. DETAILED DESCRIPTION

[0030] Now, various exemplary embodiments of the present invention are described in detail, and this detailed description should not be considered as a limitation of the present invention, but should be understood as a more detailed description of certain aspects, characteristics and embodiments of the present invention. It should be understood that the terms described in the present invention are only for describing specific embodiments and are not used to limit the present invention.

[0031] In addition, for the numerical range in the present invention, it is understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0032] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0033] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0034] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0035] In the present invention, the room temperature refers to 25±2°C.

[0036] In the following examples, the method for preparing the wheat bran enzymatic hydrolysis peptide comprises the following steps:

[0037] 1) Using wheat bran as raw material, extract wheat bran protein by alkali dissolution and acid precipitation method:

[0038] The wheat bran was crushed and passed through a 60-mesh sieve, and defatted with n-hexane to obtain defatted wheat bran;

[0039] According to the solid-liquid ratio of defatted wheat bran to water being 1:10, defatted wheat bran was added to water, and the pH of the reaction system was adjusted to 10.0 with 1 mol / L NaOH, stirred at 50°C for 1 hour, and centrifuged at 4500 rpm for 10 minutes to obtain a supernatant;

[0040] The pH of the supernatant was adjusted to 4.0 with 1 mol / L HCl, and the precipitate was recovered. Then, the pH of the supernatant was adjusted to 7.0 with 1 mol / L NaOH, and the supernatant was frozen to obtain wheat gluten protein.

[0041] 2) Using a composite protease to enzymatically hydrolyze the wheat gluten protein to obtain the wheat gluten enzymatic hydrolyzed peptide:

[0042] According to the mass volume ratio of gluten protein to water being 2g:100mL, the gluten protein obtained in step 1) is dissolved in water, and a composite protease is added at an enzyme amount of 14000U / g, and hydrolyzed for 3h at 45°C and pH=7.8, and then heated at 95°C for 10min to inactivate the composite protease, thereby obtaining a gluten protein hydrolyzate;

[0043] The amount of the wheat bran protein hydrolysate added is controlled to be 2 / 3 of the volume of the dialysis bag, and the wheat bran protein hydrolysate is added to the dialysis bag washed with distilled water, and the bag is sealed;

[0044] The dialysis bag containing the wheat bran protein hydrolysate was placed in distilled water at 4°C and magnetically stirred for 24 hours to obtain the desalted wheat bran protein hydrolysate;

[0045] The desalted wheat bran enzymatic hydrolysate was first placed in a -20°C refrigerator for 12 hours, then placed in a -80°C refrigerator for 24 hours, and finally freeze-dried for 72 hours to obtain the wheat bran enzymatic hydrolysate peptide.

[0046] The composite protease of the present invention contains Bacillus subtilis, and its enzyme activity is 120U / mg.

[0047] In the following examples, the method for preparing wolfberry juice comprises the following steps:

[0048] Soaking wolfberries in distilled water at room temperature for 4 hours, then mixing the soaked wolfberries with water at a mass volume ratio of 1 g: 2 mL, and beating to obtain the wolfberry juice;

[0049] In the following examples, a method for preparing a wheat gluten enzymatic peptide / wolfberry composite beverage comprises the following steps:

[0050] 1) Weigh each raw material according to the following mass percentage:

[0051] Wheat gluten enzymatic peptide 15%-35%, wolfberry juice 1.4%-2.2%, stevioside 4%-8%, citric acid 0.2%-0.4%, sodium carboxymethyl cellulose 0.1%, xanthan gum 0.08%, balance water;

[0052] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0053] Examples 1 to 5

[0054] A wheat bran enzymatic peptide / wolfberry compound beverage

[0055] 1) Weigh each raw material according to the mass percentage recorded in Table 1;

[0056] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0057] Table 1 Mass percentage of each raw material

[0058]

[0059] The bran enzymatic peptide / wolfberry composite beverages prepared in Examples 1 to 5 were subjected to sensory evaluation. Specifically, the sensory evaluation team consisted of 16 food major students, and each sample was scored with reference to the sensory evaluation indexes and scoring criteria. Each sample was subjected to three sensory evaluations, and the average score was calculated. The sensory evaluation indexes and scoring criteria are shown in Table 2. The sensory evaluation results are shown in Table 2. Figure 1 As shown in the figure, different letters represent significant differences at P < 0.05;

[0060] Table 2 Sensory evaluation indicators and scoring criteria

[0061]

[0062]

[0063] Depend on Figure 1 It can be seen that when the addition amount of the wheat gluten enzymatic peptides is 30%, the wheat gluten enzymatic peptides / wolfberry composite beverage prepared by the present invention has the highest sensory evaluation score (P<0.05), and at this time, the wheat gluten enzymatic peptides / wolfberry composite beverage prepared by the present invention has the unique fragrance of wheat bran; when the addition amount of the wheat gluten enzymatic peptides is greater than 30%, the sensory score decreases, because the taste of the wheat bran is too strong and masks the taste of the wolfberry; and when the addition amount of the wheat gluten enzymatic peptides is less than 30%, the sensory score decreases, because the wolfberry taste masks the taste of the wheat bran; therefore, the addition amount of the wheat gluten enzymatic peptides is preferably 30%.

[0064] Embodiments 6 to 10

[0065] A wheat bran enzymatic peptide / wolfberry compound beverage

[0066] 1) Weigh each raw material according to the mass percentage recorded in Table 3;

[0067] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0068] Table 3 Mass percentage of each raw material

[0069]

[0070]

[0071] The bran enzymatic peptide / wolfberry composite beverages prepared in Examples 6 to 10 were subjected to sensory evaluation. Specifically, the sensory evaluation team consisted of 16 food major students, and each sample was scored with reference to the sensory evaluation indexes and scoring criteria. Each sample was subjected to three sensory evaluations, and the average score was calculated. The sensory evaluation indexes and scoring criteria are shown in Table 2. The sensory evaluation results are shown in Table 2. Figure 2 As shown in the figure, different letters represent significant differences at P < 0.05;

[0072] Depend on Figure 2 It can be seen that when the addition amount of wolfberry juice is 2%, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the highest sensory evaluation score (P<0.05). At this time, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the best taste and good acidity and sweetness; when the addition amount of wolfberry juice is greater than 2.0%, the sensory score decreases, because excessive wolfberry juice makes the composite beverage taste bitter, and the wolfberry taste is too strong, which covers up the fragrant taste of wheat bran; when the addition amount of wolfberry juice is less than 2.0%, the sensory score decreases, because the taste of wheat bran is too strong; therefore, the addition amount of wolfberry juice is preferably 2.0%.

[0073] Examples 11 to 15

[0074] A wheat bran enzymatic peptide / wolfberry compound beverage

[0075] 1) Weigh each raw material according to the mass percentage recorded in Table 4;

[0076] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0077] Table 4 Mass percentage of each raw material

[0078]

[0079]

[0080] The bran enzymatic peptide / wolfberry composite beverages prepared in Examples 11 to 15 were subjected to sensory evaluation. Specifically, the sensory evaluation team consisted of 16 food major students, and each sample was scored with reference to the sensory evaluation indexes and scoring criteria. Each sample was subjected to three sensory evaluations, and the average score was calculated. The sensory evaluation indexes and scoring criteria are shown in Table 2. The sensory evaluation results are shown in Table 2. Figure 3 As shown in the figure, different letters represent significant differences at P < 0.05;

[0081] Depend on Figure 3 It can be seen that when the addition amount of stevioside is 6%, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the highest sensory evaluation score (P<0.05). At this time, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the best taste, coordinated sour, sweet and bitter tastes, and a full taste, and can give full play to the fragrance of wheat bran and wolfberry; when the addition amount of stevioside is greater than 6%, the sensory score decreases, because excessive addition of stevioside leads to excessive sweetness, and when the amount of drinking is large, the mouthfeel is sweet and greasy; when the addition amount of stevioside is less than 6%, the sensory score decreases, because the addition amount of stevioside is too low, resulting in too light taste and no pleasant feeling; therefore, the addition amount of stevioside is preferably 6%.

[0082] Embodiments 16 to 20

[0083] A wheat bran enzymatic peptide / wolfberry compound beverage

[0084] 1) Weigh each raw material according to the mass percentage recorded in Table 5;

[0085] 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

[0086] Table 5 Mass percentage of each raw material

[0087]

[0088]

[0089] The bran enzymatic peptide / wolfberry composite beverages prepared in Examples 16 to 20 were subjected to sensory evaluation. Specifically, the sensory evaluation team consisted of 16 food major students, and each sample was scored with reference to the sensory evaluation indexes and scoring criteria. Each sample was subjected to three sensory evaluations, and the average score was calculated. The sensory evaluation indexes and scoring criteria are shown in Table 2. The sensory evaluation results are shown in Table 2. Figure 4 As shown in the figure, different letters represent significant differences at P < 0.05;

[0090] Depend on Figure 4 It can be seen that when the addition amount of citric acid is 0.35%, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the highest sensory evaluation score (P<0.05), at this time, the bran enzymatic peptide / wolfberry composite beverage prepared by the present invention has the best flavor and has good sweetness and acidity; when the addition amount of citric acid is greater than 0.35%, the sensory score decreases, because excessive addition of citric acid leads to high acidity, which has an adverse effect on the taste of the composite beverage; when the addition amount of citric acid is less than 0.35%, the sensory score decreases, because the addition amount of citric acid is too low, the sweetness is heavy, the mouthfeel is too sweet and greasy, and an unpleasant sweetness is produced; therefore, the addition amount of citric acid is preferably 0.35%.

[0091] In order to further optimize the amount of each raw material in the wheat gluten enzymatic peptide / wolfberry composite beverage provided by the present invention, on the basis of the above single factor experimental results, an orthogonal experiment was designed to conduct a four-factor three-level orthogonal experiment on the addition amount of wheat gluten enzymatic peptide, the addition amount of wolfberry juice, the addition amount of stevioside, and the addition amount of citric acid. The orthogonal factor level table is shown in Table 6;

[0092] Table 6 Orthogonal factor level table

[0093]

[0094] The orthogonal test results are shown in Table 7;

[0095] Table 7 Orthogonal test results

[0096]

[0097] From the K value in the data of Table 7, it can be seen that the optimal raw material dosage of the gluten enzymatic peptide / wolfberry composite beverage provided by the present invention is: A1B2C2D2, that is, the addition amount of the gluten enzymatic peptide is 30%, the addition amount of wolfberry juice is 2%, the addition amount of stevioside is 5%, and the addition amount of citric acid is 0.35%; from the range value (R) in the data of Table 7, it can be seen that the factors of the raw materials in the gluten enzymatic peptide / wolfberry composite beverage provided by the present invention are: D>A>B>C, that is, the addition amount of citric acid>the addition amount of stevioside>the addition amount of the gluten enzymatic peptide>the addition amount of wolfberry juice;

[0098] The variance analysis of the orthogonal test results is shown in Table 8;

[0099] Table 8 ANOVA of orthogonal test results

[0100]

[0101]

[0102] Note: "*" indicates significant difference, P < 0.05; "**" indicates extremely significant difference, P < 0.01;

[0103] From the data in Table 8, it can be seen that the addition amount of stevia and citric acid has an extremely significant effect on the sensory score (P < 0.01), and the addition amount of gluten enzymatic peptide has a significant effect on the sensory score (P < 0.05). The order of factors of each raw material in the gluten enzymatic peptide / wolfberry compound beverage is: D>A>B>C, that is, the addition amount of citric acid>the addition amount of stevia>the addition amount of gluten enzymatic peptide>the addition amount of wolfberry juice, which is consistent with the results of the range analysis;

[0104] The orthogonal optimization conditions were repeatedly verified, and the raw material dosage of A1B2C2D2 was obtained, and the sensory evaluation was 93.50±1.54, which was better than other experimental groups and had the highest sensory evaluation.

[0105] The gluten enzymatic peptide / wolfberry composite beverage was prepared with the above-optimized amounts of each raw material, and the physical and chemical indicators of the gluten enzymatic peptide / wolfberry composite beverage were detected. The specific detection method was: the pH value of the gluten enzymatic peptide / wolfberry composite beverage was detected using a pH meter. After detection, the pH value of the gluten enzymatic peptide / wolfberry composite beverage prepared by the present invention was 3.99±0.02.

[0106] The wheat gluten enzymatic peptide / wolfberry composite beverage was prepared with the above optimized amounts of each raw material, and the in vitro lipid-lowering activity of the wheat gluten enzymatic peptide / wolfberry composite beverage was tested. The specific testing method is as follows:

[0107] Determination of the inhibition rate of cholesterol micelle solubility:

[0108] Preparation of cholesterol micelle solution:

[0109] ① Add 160 μL 0.1 mol / L cholesterol, 80 μL 0.05 mol / L monooleylglyceride, 80 μL 0.1 mol / L oleic acid, and 80 μL 0.06 mol / L L-α-phosphatidylcholine to a 50 mL centrifuge tube, stir for 30 seconds, and use N2 to evaporate the organic solvent in the solution;

[0110] ② Add 28.39 mg 6.6 mmol / L taurocholic acid and 8 mL 15 mmol / L phosphate buffer to a 50 mL centrifuge tube and stir to dissolve;

[0111] Pour ② into ①, stir for 2 min, treat with ultrasound (power 25%) for 3 min, incubate in a constant temperature shaking water bath (37°C, 24 h, 140 rpm), and stir;

[0112] The sample solution was mixed with the cholesterol micelle solution, stirred for 2 minutes, ultrasonically treated (power of 25%) for 3 minutes, incubated in a constant temperature shaking water bath (37°C, 140rpm, oscillation for 2 hours), and the centrifuge tube was sealed with a sealing film; after stirring, it was filtered through a 0.22μm water-soluble filter membrane, and the supernatant was collected and the cholesterol content in the micelle was determined using a total cholesterol kit, and the absorbance value was determined at 510nm to calculate the cholesterol content in the supernatant. Cholestyramine was used as a positive control, and all measurements were repeated four times in parallel, and the inhibition rate was calculated according to the following formula;

[0113]

[0114] Where:

[0115] S0-cholesterol mass concentration of blank group, mmol / L;

[0116] S1-Cholesterol mass concentration of sample group, mmol / L;

[0117] Determination of cholesterol esterase inhibition rate:

[0118] Prepare 0.1 mol / L NaCl solution, 0.2 mmol / L P-NPB solution, 5.16 mmol / L sodium taurocholate solution, 0.1 mol / L sodium phosphate buffer (pH=7.04), sample solution, 0.72 U / mL cholesterol esterase solution; use simvastatin as a positive control, refer to the reaction system described in Table 9, and perform three parallel determinations;

[0119] Table 9 Reaction system

[0120]

[0121] The inhibition rate was calculated according to the following formula;

[0122]

[0123] Where:

[0124] A-absorbance value of the original enzyme solution tube;

[0125] B-absorbance value of blank tube;

[0126] C-absorbance value of the sample experimental tube;

[0127] D- Absorbance value of the sample control tube.

[0128] Determination of pancreatic lipase inhibition rate:

[0129] Weigh 10 mg of pancreatic lipase, dissolve it in 10 mL of Tris-HCl (pH=8.0) buffer, shake well to obtain a 1.0 mg / mL PL solution; accurately transfer 8.44 μL of P-NPB solution into a 10 mL centrifuge tube, add 4 mL of acetonitrile to obtain a 12 mmol / L P-NPB solution;

[0130] PL activity inhibition rate detection:

[0131] AE: Accurately pipette 200 μL PL and 1150 μL Tris-HCl into a 1.5 mL centrifuge tube and shake thoroughly;

[0132] AIB: Accurately pipette 400 μL of sample and 600 μL of Tris-HCl into a 1.5 mL centrifuge tube and shake thoroughly;

[0133] AI: In a 2.0 mL centrifuge tube, accurately pipette 400 μL of sample, 200 μL of PL, and 1150 μL of Tris-HCl and shake thoroughly;

[0134] Pipette 200 μL of AIB into a 96-well plate; Pipette 175 μL of AE and AI into a 96-well plate respectively, and then add 25 μL of P-NPB solution; Set up 4 replicate wells for each solution and incubate at 37°C and 300 rpm for 25 min; Use an ELISA reader to measure and test, with a measurement wavelength of 405 nm; Use rutin as a positive control, measure four times in parallel, and calculate the inhibition rate according to the following formula;

[0135]

[0136] Where:

[0137] Absorbance value of AE-original enzyme solution tube;

[0138] AI-absorbance value of sample experimental tube;

[0139] AIB-absorbance value of sample control tube;

[0140] The results of in vitro hypolipidemic activity test of wheat bran proteolytic peptide / wolfberry compound beverage are as follows Figure 5 As shown, A is the cholesterol micelle solubility inhibition rate, B is the cholesterol esterase inhibition rate, and C is the pancreatic lipase inhibition rate;

[0141] Depend on Figure 5 From Figure A, we can see that the inhibition rate of cholesterol micelle solubility of wheat gluten enzymatic peptide / wolfberry composite beverage was 81.44±3.30%, which was lower than the positive control cholestyramine, but significantly higher than that of wheat gluten enzymatic peptide or wolfberry juice (P<0.05);

[0142] Depend on Figure 5From Figure B, we can see that the cholesterol esterase inhibition rate of the wheat gluten enzymatic peptide / wolfberry compound beverage was 57.47±2.89%, which was lower than the positive control simvastatin, slightly higher than wolfberry juice, but significantly higher than the in vitro lipid-lowering ability of the wheat gluten enzymatic peptide (P<0.05);

[0143] Depend on Figure 5 From C, we can see that the pancreatic lipase inhibition rate of the wheat gluten enzymatic peptide / wolfberry compound beverage was 58.04±3.47%, which was lower than the positive control rutin, slightly higher than wolfberry juice, but significantly higher than the in vitro lipid-lowering ability of the wheat gluten enzymatic peptide (P<0.05);

[0144] Therefore, the wheat gluten enzymatic hydrolyzed peptide / wolfberry composite beverage provided by the present invention has good in vitro blood lipid lowering activity.

[0145] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the above embodiments, ordinary technicians in the relevant field should understand that the specific implementation methods of the present invention can still be modified or replaced by equivalents. Any modification or equivalent replacement that does not depart from the spirit and scope of the present invention should be covered within the scope of protection of the claims of the present invention.

Claims

1. A wheat gluten enzymatic peptide / wolfberry composite beverage, characterized in that: Including the following raw materials in mass percentage: 15% to 35% of wheat gluten enzymatic peptides, 1.4% to 2.2% of wolfberry juice, 4% to 8% of stevioside, 0.2% to 0.4% of citric acid, 0.1% of sodium carboxymethyl cellulose, 0.08% of xanthan gum, and the balance of water.

2. A wheat gluten enzymatic peptide / wolfberry composite beverage according to claim 1, characterized in that: The method for preparing the wheat bran enzymatic hydrolysis peptide comprises the following steps: 1) Using wheat bran as raw material, extracting wheat bran protein by alkali dissolution and acid precipitation method; 2) Using a composite protease to enzymatically hydrolyze the gluten protein obtained in step 1) to obtain the gluten enzymatic hydrolyzed peptide.

3. A wheat gluten enzymatic peptide / wolfberry composite beverage according to claim 2, characterized in that: In step 1), the alkali dissolution and acid precipitation method is specifically as follows: the wheat bran is crushed through a 60-mesh sieve and defatted with n-hexane to obtain defatted wheat bran; the defatted wheat bran is added to water according to a solid-liquid ratio of defatted wheat bran to water of 1:10, and the pH of the reaction system is adjusted to 10.0 with 1 mol / L NaOH, stirred at 50°C for 1 hour, and centrifuged at 4500 rpm for 10 minutes to obtain a supernatant; the pH of the supernatant is adjusted to 4.0 with 1 mol / L HCl, the precipitate is recovered, and then the pH of the supernatant is adjusted to 7.0 with 1 mol / L NaOH, and frozen to obtain the wheat bran protein.

4. A wheat gluten enzymatic peptide / wolfberry composite beverage according to claim 2, characterized in that: In step 2), the enzymolysis is specifically as follows: dissolving the gluten protein obtained in step 1) in water according to a mass volume ratio of gluten protein to water of 2g:100mL, adding a composite protease according to an enzyme addition amount of 14000U / g, hydrolyzing for 3h at 45°C and pH=7.8, and then heating at 95°C for 10min to inactivate the composite protease to obtain a gluten protein hydrolysate; controlling the amount of the gluten protein hydrolysate added to be 2 / 3 of the volume of the dialysis bag, adding the gluten protein hydrolysate to the dialysis bag washed with distilled water, and sealing the bag; placing the dialysis bag containing the gluten protein hydrolysate in 4°C distilled water, stirring magnetically for 24h, and obtaining a desalted gluten protein hydrolysate; first placing the desalted gluten protein hydrolysate in a -20°C refrigerator, freezing for 12h, then placing it in a -80°C refrigerator, freezing for 24h, and finally freeze-drying for 72h.

5. A wheat gluten enzymatic peptide / wolfberry composite beverage according to claim 4, characterized in that: The composite protease contains Bacillus subtilis, and its enzyme activity is 120U / mg.

6. A wheat gluten enzymatic peptide / wolfberry composite beverage according to claim 1, characterized in that: The method for preparing wolfberry juice comprises the following steps: The wolfberries were soaked in distilled water at room temperature for 4 hours, and then the soaked wolfberries were mixed with water at a mass volume ratio of 1 g: 2 mL, and pulped to obtain the wolfberry juice.

7. A method for preparing the wheat gluten enzymatic peptide / wolfberry composite beverage as claimed in claim 1, comprising the following steps: 1) Weigh each raw material according to the above mass percentage; 2) The raw materials weighed in step 1) are mixed, homogenized at 20 MPa for 5 minutes, filled and sealed, and then pasteurized at 65° C. for 30 minutes, and cooled to room temperature to obtain the wheat gluten enzymatic peptide / wolfberry composite beverage.

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